Deposited Dose Rate and Dissolution of Nanoparticles in the Respiratory Tract Impact the Acute ...

209 Pages·2013·4.47 MB·English

Deposited Dose Rate and Dissolution of Nanoparticles in the Respiratory Tract
Impact the Acute Inflammatory Response

by
Brittany Lynn (Serke) Baisch

Submitted in Partial Fulfillment
of the
Requirements for the Degree
Doctor of Philosophy

Supervised by
Associate Professor Alison C.P. Elder

Toxicology Training Program
Department of Environmental Medicine
School of Medicine and Dentistry

University of Rochester
Rochester, NY
2013
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BIOGRAPHICAL SKETCH
Brittany Lynn (Serke) Baisch graduated summa cum laude from Western
Connecticut State University (WCSU) in 2008, with a triple baccalaureate of arts in
Chemistry, Biochemistry and Spanish. She was awarded several scholarships and
prestigious awards as an undergraduate, including the Dean of Arts & Sciences Award,
the WCSU/American Chemical Society General Chemistry Student of the Year Award
and Outstanding Senior of the Year Award. She also received the Henry Barnard
Outstanding Student Award along with an Official Citation from the State of Connecticut
Legislature for Outstanding Academic Achievement and Community Service. Prior to
graduate school, Brittany worked as a formulations chemist at Advanced Technology
Materials, Inc. where she converted cleaning chemistries used for logic and
microelectronic devices to green formulations. Her work resulted in the co-authorship of
two patents:
Sonthalia P, Cooper E, Minsek D, Zhang P, Petruska M, Serke BL, Hurd T. Non-
selective oxide etch wet clean composition and method of use. World Intellectual
Property Organization WO/2009/111719, International Patent, Nov 2009.
Korzenski MB, Jiang P, Serke BL. Non-fluoride containing composition for the
removal of residue from a microelectronic device. World Intellectual Property
Organization WO/2009/032460, International Patent, Dec 2009.

Brittany joined the Toxicology Training Program at the University of Rochester
in 2008 and the laboratory of Dr. Alison Elder in 2009, where she studied the role of
deposited dose rate in nanoparticle-induced inflammatory responses in respiratory tract
target cells. She successfully passed her Qualifying Exam for her M.S. in Toxicology in
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2011. Brittany has earned several awards for her research, including an internship at
Bristol-Myers Squibb (2009), Colt Foundation Travel Award for the Nanotoxicology
Conference in Edinburgh, Scotland (2010), 1st Place Outstanding Graduate Student
Award from the Society of Toxicology (SOT)’s Nanotoxicology Specialty Section
(2011), the SOT Graduate Student Travel Award (2012), Outstanding Presentation
Award at the Nanotoxicology Conference in Beijing, China (2012), and 2nd place at the
University of Rochester’s Graduate Student Society Poster session in the
Multicellular/Organismal Category (2013). She has been a member of SOT since 2009
and belongs to its Nanotoxicology and Inhalation and Respiratory Specialty Sections. The
following peer-reviewed publications summarize her graduate work, with 2 additional
manuscripts in preparation:
Baer DR, Engelhard M, Johnson G, Laskin J, Mueller K, Munusamy P,
Thevuthasan S, Wang H, Washton N, Elder A, Baisch BL, Karakoti A, Kuchibhatla SV,
Moon D. Surface Characterization of Nanomaterials: Important Needs and Challenging
Opportunities. Journal of Vacuum Science and Technology A. In press.

Baisch BL, Corson NM, Wade-Mercer P, Gelein R, Kennell AJ, Oberdörster G,
Elder A. Equivalent titanium dioxide nanoparticle deposition by intratracheal instillation
and whole body inhalation: The effect of dose rate on acute respiratory tract
inflammation. Particle and Fibre Toxicology. Submitted.

She is the current student representative for SOT’s Committee on Diversity
Initiatives and the Women in Toxicology Special Interest Group. Brittany is also a
Docent at the Seneca Park Zoo (2010-present) and volunteers as a Science Ambassador
to teach laboratory-oriented case studies in the Rochester area public schools and
retirement communities (2010-present). She also held a Scientist Instructor position in the
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Life Sciences Learning Center in the Department of Environmental Medicine at the
University of Rochester (2011) and was the teaching assistant for the Biochemical
Toxicology course (2012).
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ACKNOWLEDGEMENTS
Earning the Ph.D. has been an overwhelming amount of work and dedication, but
it would not be possible without several sets of helping hands, people to guide me in the
right direction and others in my life who have made sure I have stayed on course by
always remembering what is important.
First, and most importantly, is my advisor, Alison. You have been wonderful to
work with over the years. You are just so normal, and by scientific standards that is often
times difficult to come by. I am always paranoid and stressed about everything, including
hypothetical situations that never turn into reality, and you are always calm and maintain
good rapport with everyone, whether or not they deserve it. I thank you for your calm
demeanor and for your patience while I have assembled presentations, posters and several
pieces of writing, barely making it by the deadlines. You are very detailed oriented,
which has trained me to think about more than just the “big picture.” I am not going to
lie; it has been incredibly difficult being the first, and only, graduate student in the
laboratory. It was very frustrating in the beginning to figure out what project to work on
and which avenue to pursue. It was very time consuming and overwhelming to write so
many protocols for the laboratory and to learn so much about the data sets over the years.
However, you have always been approachable and receptive to my scientific and non-
scientific thoughts and that has made all of the difference, and really, truly, why I have
stuck it out until the end! Thank you for everything and I look forward to being
colleagues in the future.
Secondly, I need to thank Günter “GO” Oberdörster. Whether you would like to
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accept it or not, you are the grandfather of nanotoxicology and you and Alison have made
amazing contributions to the field. It has always made me giggle when we go to
conferences and there is a crowd of people waiting to shake your hand and to get a
picture taken with you. Then I realize that I have had the honor of working with you
everyday and I am appreciative of how fortunate I am. Although there are times when I
think I am hilarious and funny when I poke fun at your competitive spirit and healthy
lifestyle, and times when I have annoyed you when you were on the phone or trying to
prepare for one of your “international tours,” but you have been incredibly helpful,
knowledgeable and honest every step of the way and have made time to listen to my rants
and thoughts that have often times made no sense. Thank you for your expert advice,
laughs and for all of the voicemails you have left on my phone trying to get back to me or
looking for me; whether it has been to provide feedback on my data interpretations, or to
fetch you a spoon from the break room, or to help you align text in the Prism software, it
has been a lot of fun. It has been a pleasure getting to know, and working with, “The
Great One” of the nanotoxicology field.
Thirdly, I would like to thank the other members of my thesis advisory
committee: Drs. Jack Finkelstein, Gail V.W. Johnson and Lisa DeLouise. I appreciate
your thoughts, insights, suggestions and critiques over the past few years. I have enjoyed
growing as a scientist under your guidance and it has been inspiring to witness how
interested and invested you have been in my project(s). Your curiosity and ideas have
reminded me why we have all fallen in love with science.
I am unceasingly thankful to the past and current members of the
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Oberdörster/Elder laboratory: Nancy Corson, Pam Wade-Mercer, Bob Gelein, Andrea
Walker and Judy Havalack. This paragraph does no justice in describing how truly
grateful I am for how patient you have been with my demanding personality and all of the
time you have spent helping me with my project. Whether it was training me, ordering
supplies, helping to collect samples, run samples, perform exposures and sacrifices or
assist with administrative tasks, you never complained about it and we always worked
together as a team. I was very lucky to work alongside such hard working, motivated,
dedicated, patient, kind, caring, smart and well-qualified people. We had so much fun
outside of lab with our harvest parties, birthday celebrations and walks through Mt. Hope
Cemetery. I will miss all of you and hope you will always stay in touch.
I would like to acknowledge all of our internal and external collaborators who
have been of great assistance with their helpful discussions and by sharing protocols. At
the University of Rochester I would like to thank the laboratories of Drs. Irfan Rahman,
Rick Phipps, Paige Lawrence, Michael O’Reilly and Jack Finkelstein as well as Dr. Miao
Shi for helpful discussion about NP characterization and for performing some XRD
measurements. I also appreciate all of the TEM analysis that Karen Bentley (URMC EM
Core) performed on dozens of samples over the past four years. We have collaborated
with several people at Pacific Northwest National Laboratories / Environmental
Molecular Sciences Laboratory in Richland, Washington who have assisted with the
characterization of our materials. Specifically, I would like to acknowledge the following
people and their corresponding contributions over the years: Drs. Justin Teeguarden (In
Vitro Sedimentation and Diffusion Dosimetry model calculations), Galya Orr (helpful
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discussions regarding in vitro NP deposition using ALI systems), Don Baer (for including
our laboratory in a co-authored review paper regarding the surface characterization of our
in-house generated mixed copper oxide NPs), and Mark Engelhard (XPS characterization
of our copper containing NPs). I would also like to thank Drs. Julian Taurozzi and Vince
Hackley at the National Institute of Standards and Technology, for performing the LDS
measurements on the TiO NPs. Lastly, I would like to thank Drs. Manoranjan Sahu and
2
Pratim Biswas from the University of Washington, St. Louis, for generating and helping
to characterize the suite of Cu-TiO NPs.
2
The sixth “ thank you” I have is for the Department of Environmental Medicine.
Namely, Chris Gramza and Drs. Tom Gasiewicz, Paige Lawrence, the late Ned Ballatori,
Pat Noonan-Sullivan, Cathy Amico, Deb Bendschneider, Pat Lombardo, and Laurie
Furibondo for maintaining such an outstanding and meticulous track record of the
Toxicology Training Program. I am convinced that there is no other toxicology program
in the country with such a renowned, collaborative, spirited and scientifically impressive
group of faculty and alumni. Chris has been like a mom to all of the students and has
been supportive and a true friend throughout this whole process. I am certain that I would
not have scheduled anything or turned in any paper work on time without her guidance
and initiative. I also have to thank Drs. Dina Markowitz, Liam Casey and Shaw-Ree
Chen for the science outreach and teaching opportunities I had in the Life Sciences
Learning Center, which have improved my presentation style and how I convey scientific
knowledge to the public.
I would also like to thank my fellow toxicology students, the alumni, my friends,
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as well as my former co-workers at ATMI for all of their support, helpful discussion and
career advice, especially in this last year. Without the support and feedback of my peers I
would not have made it through the program! I have made wonderful, lifelong friends
who have reminded me that nothing replaces hard work, to always stand my ground and
to give credit where credit is due.
Last, but certainly not least, I would like to thank my husband, Tom. No one else
has made greater sacrifices than you have. You have given up many things you have
wanted for yourself and for your own career so that I could have this opportunity to go to
graduate school. There were times when you worked two or three jobs so that we could
pay for our wedding and make ends meet. You have worked over 1,600 weekends,
cooked over 1,200 dinners, walked the dog over 3,000 times, and hosted our family 20
times, but you have never complained about anything. In fact, you have encouraged me
to never quit and have convinced me that I could always get it done. You made me
realize that it is okay to relax and have fun once in a while, too. I cannot wait for the next
step in our life together and I know that you have been patiently waiting for this to come
to an end for a long, five years. We can both agree that our health, our happiness, our
hope and the strength of our love for one another, has been provided only through the
grace of God. Our praise has always gone up to Him and His blessings have always fallen
down on us.
“For I know the plans I have for you, declares the Lord, plans to prosper you and
not to harm you, plans to give you hope and a future.” - Jeremiah 29:11
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ABSTRACT
The increasing use of nanoparticles (NPs) has led to concerns about the potential
for human exposures and associated health outcomes. Although there are several likely
routes of exposure to NPs, we focus on the respiratory tract (RT) and subsequent effects
in the lungs. Since there is a lack of real world exposure data for NPs, no RT exposure
limits have been imposed by regulatory agencies. Due to the overwhelming number of
NPs being produced, it is not possible to interrogate all of them for hazard assessment
purposes. Therefore, there is a need for exposure models that reflect real world exposures
and that are adequately predictive of hazard for NPs that share the same physicochemical
properties. Toxicological responses to NPs have been explored, typically using high dose
rate (bolus) delivery and high doses; however, little is known about the roles of dose rate
and dissolution rate in determining response outcomes. We hypothesize that the NP
deposited dose rate and dissolution rate are key determinants of the acute inflammatory
responses in the RT when the deposited dose is held constant. We exposed F-344 male
rats to poorly soluble titanium dioxide (TiO ), partially soluble copper-doped TiO , or
2 2
partially soluble copper oxide NPs by high dose rate intratracheal instillation or low dose
rate whole body inhalation. We observed more robust inflammation, hallmarked by
neutrophil influx and neutrophil chemoattractant proteins, following high dose rate
instillation of poorly soluble TiO NPs compared to low dose rate inhalation. Therefore,
2
high dose rate delivery of the poorly soluble TiO NPs overestimates NP hazard. Using
2
partially soluble Cu containing NPs, we demonstrated that both the deposited dose rate
and dissolution rate impact inflammation. Our studies provide a better understanding of